Ketobenzamides as calpain inhibitors

ABSTRACT

The patent describes ketobenzamides of the formula ##STR1## where R1, R2, R3, R4, X and n have the meanings given in the description, and their preparation. The novel compounds are useful for controlling disorders.

This application is a 371 of PCT/EP97/06655 Nov. 28, 1997.

The present invention relates to novel ketobenzamides, to theirpreparation and to their use in controlling diseases.

Calpains are intracellular, proteolytic enzymes from the cysteineprotease group and are found in many cells. Calpains are activated byelevated calcium concentration, with a distinction being made betweencalpain I or μ-calpain, which is activated by μ-molar concentrations ofcalcium ions, and calpain II or m-calpain, which is activated by m-molarconcentrations of calcium ions (P. Johnson, Int. J. Biochem. 1990,22(8), 811-22). Further calpain isoenzymes are currently beingpostulated (K. Suzuki et al., Biol. Chem. Hoppe-Seyler, 1995,376(9),523-9).

It is assumed that calpains play an important role in variousphysiological processes. These processes include the cleavage ofregulatory proteins such as protein kinase C, cytoskeletal proteins suchas MAP 2 and spectrin, and muscle proteins, protein degradation inrheumatoid arthritis, proteins in the activation of platelets,neuropeptide metabolism, proteins in mitosis, and additional exampleswhich are listed in M. J. Barrett et al., Life Sci. 1991, 48, 1659-69and K. K. Wang et al., Trends in Pharmacol. Sci., 1994, 15, 412-9.

Elevated calpain levels can be measured in various pathophysiologicalprocesses, for example: ischemias of the heart (eg. myocardialinfarction), the kidney or the central nervous system (eg. stroke),inflammations, muscular dystrophies, cataracts of the eyes, injuries tothe central nervous system (eg. trauma), Alzheimer's disease, etc. (seeK. K. Wang, above). It is assumed that there is a connection betweenthese diseases and persistently elevated intracellular calcium levels.This results in calcium-dependent processes becoming hyperactivated andno longer being subject to physiological control. A correspondinghyperactivation of calpains can also trigger pathophysiologicalprocesses.

For this reason, it was postulated that inhibitors of the calpainenzymes could be of use for treating these diseases. This postulate isconfirmed by a variety of investigations. Thus, Seung-Chyul Hong et al.,Stroke 1994, 25(3), 663-9 and R. T. Bartus et al., Neurological Res.1995, 17, 249-58 have demonstrated that calpain inhibitors have aneuroprotective effect in acute neurodegenerative disturbances orischemias such as occur after cerebral stroke. Following experimentalbrain traumas, calpain inhibitors improved recovery from the memoryperformance deficits and neuromotor disturbances which occurred (K. E.Saatman et al. Proc. Natl. Acad. Sci. USA, 1996, 93,3428-3433). C. L.Edelstein et al., Proc.Natl.Acad.Sci. USA, 1995, 92, 7662-6, found thatcalpain inhibitors have a protective effect on hypoxia-damaged kidneys.Yoshida, Ken Ischi et al., Jap. Circ. J. 1995, 59(1), 40-8 pointed outthat calpain inhibitors had favorable effects following cardiac damagewhich was produced by ischemia or reperfusion. Since calpain inhibitorsinhibit the release of the β-AP4 protein, it was suggested that they hada potential use as therapeutic agents in Alzheimer's disease (J. Higakiet al., Neuron, 1995, 14, 651-59). The release of interleukin-1α is alsoinhibited by calpain inhibitors (N. Watanabe et al., Cytokine 1994,6(6), 597-601). It has furthermore been found that calpain inhibitorshave cytotoxic effects on tumor cells (E. Shiba et al. 20th Meeting Int.Ass. Breast Cancer Res., Sendai J p, Sep. 25-28, 1994, Int. J. Oncol.5(Suppl.), 1994, 381).

Further possible uses of calpain inhibitors are listed in K. K. Wang,Trends in Pharmacol. Sci., 1994, 15, 412-8.

Calpain inhibitors have already been described in the literature.However, these are predominantly either irreversible inhibitors orpeptide inhibitors. As a rule, irreversible inhibitors are alkylatingsubstances and suffer from the disadvantage that they reactunselectively in the organism or are unstable. Thus, these inhibitorsoften exhibit undesirable side-effects, such as toxicity, and as aresult are restricted in use or not usable at all. The epoxides E 64 (E.B. McGowan et al., Biochem. Biophys. Res. Commun. 1989, 158, 432-5),α-halo ketones (H. Angliker et al., J. Med. Chem. 1992, 35, 216-20) anddisulfides (R. Matsueda et al., Chem. Lett. 1990, 191-194), for example,can be included among the irreversible inhibitors.

Many known reversible inhibitors of cysteine proteases such as calpainare peptide aldehydes, in particular dipeptide or tripepide aldehydessuch as Z-Val-Phe-H (MDL 28170) (S. Mehdi, Trends in Biol. Sci. 1991,16, 150-3) and the compounds from EP 520336. Under physiologicalconditions, peptide aldehydes frequently suffer from the disadvantagethat they are unstable as a result of their high level of reactivity,can be rapidly metabolized and are prone to nonspecific reactions whichcan be the cause of toxic effects (J. A. Fehrentz and B. Castro,Synthesis 1983, 676-78). Consequently, peptide aldehydes are either ofonly limited usefulness, or of no use at all, in the treatment ofdiseases.

The discovery that certain peptide ketone derivatives are alsoinhibitors of cysteine proteases and calpain, in particular, representsa step forward. Thus, ketone derivatives in which the keto group isactivated by an electron-withdrawing group such as CF₃ are known to beinhibitors in the case of serine proteases, for example. Derivativeshaving ketones which are activated by CF₃ or similar groups are onlyslightly effective, or not effective at all, in the case of cysteineproteases (M. R. Angelastro et al., J. Med. Chem. 1990,33, 11-13).Surprisingly, only ketone derivatives in which, on the one hand,α-terminal leaving groups cause an irreversible inhibition and, on theother hand, the keto group is activated by a carboxylic acid derivative,have hitherto been found to be effective inhibitors in the case ofcalpain (see M. R. Angelastro et al., see above; WO 92/11850; WO92,12140; WO 94/00095 and WO 95/00535). However, it is only peptidederivatives of these keto amides and keto esters which have hithertobeen reported to be effective (Zhao Zhao Li et al., J. Med. Chem. 1993,36, 3472-80; S. L. Harbenson et al., J. Med. Chem. 1994, 37, 2918-29 andsee M. R. Angelastro et al. above).

In addition, ketobenzamides are known from the literature. Thus, theketo ester PhCO-Abu-COOCH₂ CH₃ has been described in WO 91/09801, WO94/00095 and 92/11850. However, M. R. Angelastro et al., J. Med. Chem.1990,33, 11-13 found the analogous phenyl derivative Ph-CONH--CH(CH₂Ph)--CO--COOCH₃ to be only a weak inhibitor of calpain. This derivativeis also described in J. P. Burkhardt, Tetrahedron Lett., 1988, 3433-36.However the importance of the substituted benzamides has never beeninvestigated to date.

Substituted, non-peptide ketobenzamide derivatives having an improvedeffect have now been found.

The present invention relates to ketobenzamides of the formula I##STR2## and their tautomeric and isomeric forms, and also, whereappropriate, their physiologically tolerated salts, where the variableshave the following meanings:

R¹ is phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyrazyl,pyridazyl, quinazolyl, quinoxalyl, thienyl, benzothienyl, benzofuryl,benzimidazolyl, furanyl, indolyl, isoquinoline, tetrahydroisoquinolineor tetrahydroquinoline, where the aromatic and heteroaromatic rings canadditionally be substituted by one, two or three R⁵ radicals,

R² is chlorine, bromine, fluorine, C₁ -C₆ -alkyl, C₂ -C₆ -alkenyl, C₂-C₆ -alkynyl, C₁ -C₆ -alkyl-phenyl, C₂ -C₆ -alkenyl-phenyl, C₂ -C₆-alkynyl-phenyl, phenyl, NHCO--C₁ -C₄ -alkyl, --NHCO-phenyl,--NHCO-naphthyl, H₂ N--SO₂ --C₁₋₄ -alkyl--, COOH, --COO--C₁₋₄ -alkyl,--CONH--C₁₋₄ -alkyl, C₁₋₄ -alkoxy, NO₂ or NH₂,

R³ is C₁ -C₆ -alkyl which can also carry a phenyl, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, indolyl, pyridyl ornaphthyl ring which, for its part, can be substituted by one or two R⁵radicals,

X is a bond, --(CH₂)_(m) --, --(CH₂)_(m) --O--(CH₂)_(o) --, --(CH₂)_(n)--S--(CH₂)_(m) --, --(CH₂)_(n) --SO--(CH₂)_(m) --, --(CH₂)_(n) --SO₂--(CH₂)_(m) --, --CH═CH--, --C.tbd.C--, --CO--CH═CH--, CO--(CH₂)_(m) --,--(CH₂)_(m) --NHCO--(CH₂)_(o) --, --(CH₂)_(m) --CONH--(CH₂)_(o) --,--(CH₂)_(m) --NHSO₂ --(CH₂)_(o) --, --NH--CO--CH═CH--,--CH═CH--CO--NH--, --(CH₂)_(m) --SO₂ NH--(CH₂)_(o) -- or ##STR3## R⁴ isOR⁶, NR⁷ R⁸, ##STR4## R⁵ is hydrogen, C₁ -C₄ -alkyl, --O--C₁ -C₄ -alkyl,OH, Cl, F, Br, I, CF₃, NO₂, NH₂, CN, COOH, COO--C₁ -C₄ -alkyl,--NHCO--C₁ -C₄ -alkyl, --NHCO-phenyl, --NHSO₂ --C₁ -C₄ -alkyl, --NHSO₂-phenyl, --SO₂ --C₁ -C₄ -alkyl or --SO₂ -phenyl,

R⁶ is hydrogen or C₁ -C₆ -alkyl which can be substituted by a phenylring which, itself, can also be substituted by one or two R⁹ radicals,

R⁷ is hydrogen or C₁ -C₆ -alkyl,

R⁸ is hydrogen or C₁ -C₆ -alkyl which can also be substituted by aphenyl ring, which can carry one or two R⁹ radicals, or by one of theradicals ##STR5## R⁹ is hydrogen, C₁ -C₄ -alkyl, --O--C₁ -C₄ -alkyl, OH,Cl, F, Br, I, CF₃, NO₂, NH₂, CN, COOH, COO--C₁ -C₄ -alkyl, --NHCO--C₁-C₄ -alkyl, --NHCO-phenyl, --NHSO₂ --C₁ -C₄ -alkyl, --NHSO₂ -phenyl,--SO₂ --C₁ -C₄ -alkyl or --SO₂ -phenyl,

R¹⁰ is hydrogen or C₁ -C₆ -alkyl which can be substituted by a phenylring which can also be substituted by one or two R⁹ radicals,

n is the number 0, 1 or 2,

m is the number 0, 1, 2, 3 or 4, and

o is the number 0, 1, 2, 3 or 4.

Preference is given to compounds of the formula I where

R² is hydrogen, C₁ -C₄ -alkyl, fluorine or chlorine,

R³ is --CH₂ -phenyl, --CH₂ -cyclohexyl, n-butanyl or n-pentanyl, each ofwhich can be substituted by an R⁵ radical,

R⁴ is --NR⁸, and

R¹, X and n have the meanings given in claim 1.

The compounds of the formula I can be employed as racemates or asenantiomerically pure compounds or as diastereomers. If enantiomericallypure compounds are desired, these can be obtained, for example, bycarrying out a conventional racemate resolution on the compounds of theformula I, or their intermediates, using a suitable optically activebase or acid. The enantiomeric compounds may also be prepared byemploying commercially obtainable compounds, for example opticallyactive amino acids such as phenylalanine, tryptophan and tyrosine.

The invention also relates to compounds which are mesomeric ortautomeric in relation to compounds of the formula I, for example thosecompounds in which the keto group of the formula I is present as an enoltautomer.

Some of the novel compounds I can contain a basic or an acidic group. Inthese cases, the compounds I can be present in the form of theirphysiologically tolerated salts, which can be obtained by reacting thecompounds I with a suitable acid or base.

Examples of suitable acids for forming salts with novel compounds Iwhich contain a basic group are hydrochloric acid, citric acid, tartaricacid, lactic acid, phosphoric acid, methanesulphonic acid, acetic acid,formic acid, maleic acid, fumaric acid, malic acid, succinic acid,malonic acid and sulphuric acid. Examples of suitable bases arepotassium hydroxide, sodium hydroxide, lithium hydroxide, triethylamine,α,α,α-tris(hydroxymethyl)methylamine and other amines.

The ketobenzamides I according to the invention can be prepared bydifferent routes, which are outlined in synthesis schemes 1 and 2.

The carboxylic esters II are converted into the acids III using acids orbases, such as hydrochloric acid, lithium hydroxide, sodium hydroxide orpotassium hydroxide, in aqueous medium or in mixtures of water andorganic solvents, such as alcohols or tetrahydrofuran, at roomtemperature or at elevated temperatures, such as 25-100° C. The acidsIII are linked to an α-amino acid derivative using the customaryconditions, which are listed, for example, in Houben-Weyl, Methoden derorganischen Chemie [Methods of organic chemistry], 4th edition, E5, Ch.V, and C. R. Larock, Comprehensive Organic Transformations, PublisherVCH, 1989, Ch. 9.

The carboxylic acid III is converted into an "activated" acid derivativeR¹ -L, where L is a leaving group such as Cl, imidazole andN-hydroxybenzotriazole, and converted into the derivative IV by reactionwith an amino acid derivative H₂ N--CHR³ --COOR. This reaction takesplace in anhydrous, inert solvent, such as methylene chloride,tetrahydrofuran and dimethylformamide, at from -20 to +25° C. ##STR6##

The derivatives IV, which are esters as a rule, are converted into theketo carboxylic acids V in analogy with the hydrolysis described above.The keto esters I' are prepared in a Dakin-West-analogous reaction usinga method described by Zhao Zhao Li et al. J. Med. Chem., 1993, 36,3472-80. In this reaction, a carboxylic acid such as V is reacted, atelevated temperature (50-100° C.) in solvents, such as tetrahydrofuran,with oxalyl chloride monoester and the resulting product is thenconverted into the keto ester I' according to the invention using bases,such as sodium ethoxide, in ethanol at 25-80° C. The keto esters I' canbe hydrolyzed, as described above, to give the keto carboxylic acidsaccording to the invention, for example.

The reaction to give the ketobenzamides I is also carried out in analogywith the method of Zhao Zhao Li et al. (see above). The keto group in I'is protected by adding 1,2-ethanedithiol in association with Lewis acidcatalysis, for example using boron trifluoride etherate, in inertsolvents, such as methylene chloride, at room temperature, resulting ina dithiane. These derivatives are reacted with amines R⁴ -H in polarsolvents, such as alcohols, at temperatures of 0-80° C., resulting inthe formation of the keto amides I (eg. R⁴ =NR⁷ R⁸). ##STR7##

An alternative method is depicted in Scheme 2. The keto carboxylic acidsIII are reacted with the amino hydroxy carboxylic acid derivatives IV(for preparation of IV, see S. L. Harbenson et al., J. Med. Chem. 1994,37, 2918-29) using the customary peptide coupling methods (seeHouben-Weyl above), resulting in the formation of the amides VII. Thesealcohol derivatives VII can be oxidized to give the keto carboxylic acidderivatives I according to the invention. A variety of customaryoxidation reactions (see C. R. Larock, Comprehensive OrganicTransformations, Publisher VCH, 1989, pages 604 f.), such as Swernoxidations and Swern-analogous oxidations, preferably using dimethylsulfoxide/pyridine-sulfur trioxide in solvents such as methylenechloride or tetrahydrofuran, with or without the addition of dimethylsulfoxide, at room temperature or at temperatures of -50 to 25° C., (T.T. Tidwell, Synthesis 1990, 857-70) or sodium hypochloride/TEMPO (S. L.Harbenson et al., see above), can be used for this purpose.

When the compounds VII are a-hydroxy esters (X=O-alkyl), these can behydrolyzed to give the carboxylic acids VIII in analogy with the abovemethods, but preferably using lithium hydroxide in water/tetrahydrofuranmixtures at room temperature. Other esters or amides X are prepared byreaction with alcohols or amines under coupling conditions which havealready been described. The alcohol derivative X can once again beoxidized to give keto carboxylic acid derivatives I according to theinvention.

The synthesis of the carboxylic esters II has already been described insome cases, or can be carried out in accordance with customary chemicalmethods.

Compounds in which X is a bond are prepared by customary aromaticcoupling, for example the Suzuki coupling using boric acid derivativesand halides, while catalyzing with palladium, or the copper-catalyzedcoupling of aromatic halides. The alkyl-bridged radicals (X=--(CH₂)_(m)--) can be prepared by reducing the analogous ketones or by alkylatingthe organolithium compounds, for example ortho-phenyloxazolidines, orother organometallic compounds (cf. I. M. Dordor, et al., J. Chem. Soc.Perkin Trans. I, 1984, 1247-52).

Ether-bridged derivatives are prepared by alkylating the correspondingalcohols or phenols using halides.

The sulfoxides and sulfones can be obtained by oxidizing thecorresponding thioethers.

Alkene-bridged and alkyne-bridged compounds are prepared, for example,from aromatic halides and corresponding alkenes and alkynes using theHeck reaction (cf. I. Sakamoto et al., Chem. Pharm. Bull., 1986, 34,2754-59).

The chalcones are produced by the condensation of acetophenones withaldehydes and can, where appropriate, be converted into the analogousalkyl derivatives by means of hydrogenation.

Amides and sulfonamides are prepared from the amines and acidderivatives in analogy with the above-described methods.

The ketobenzamides I according to the invention are inhibitors ofcysteine proteases, in particular of cysteine proteases such as calpainsI and II and cathepsins B and L.

The inhibitory effect of the ketobenzamides I was ascertained usingenzyme tests which are customary in the literature, with theconcentration of the inhibitor at which 50% of the enzyme activity isinhibited being determined as the criterion of efficacy. This approachwas used to measure the inhibitory effect of the benzamides I on calpainI, calpain II and cathepsin B.

Cathepsin B Test

The inhibition of cathepsin B was determined in analogy with a methoddescribed by S. Hasnain et al., J. Biol. Chem. 1993, 268, 235-40.

2 μL of an inhibitor solution, which was prepared from inhibitor andDMSO (final concentrations: 100 μM to 0.01 μM) were added to 88 μL ofcathepsin B (human liver cathepsin B (Calbiochem), diluted to 5 units in500 μM buffer). This mixture is preincubated at room temperature (25°C.) for 60 minutes and the reaction is then started by adding 10 μL of10 mM Z-Arg--Arg-pNA (in buffer containing 10% DMSO). The reaction ismonitored at 405 nm for 30 minutes in a microtiter plate reader. TheIC₅₀ 's are then determined from the maximum slopes.

Calpain I and Calpain II Test

The inhibitory properties of calpain inhibitors are tested in buffercontaining 50 mM tris-HCl, pH 7.5; 0.1 M NaCl; 1 mM dithiotreithol[sic]: 0.11 mM CaCl₂, using the fluorogenic calpain substrateSuc-Leu-Tyr-AMC (25 mM, dissolved in DMSO, Bachem/Switzerland) (Sasakiet al. J. Biol. Chem. 1984, Vol. 259, 12489-12494). Human μ-calpain isisolated from erythrocytes following the methods of Croall and DeMartino(BBA 1984, Vol. 788, 348-355) and Graybill et al. (Bioorg. & Med. Lett.1995, Vol. 5, 387-392). After several chromatographic steps (DEAESepharose, phenyl Sepharose, Superdex 200 and Blue Sepharose), theenzyme is obtained at a purity of <95%, as assessed by SDS-PAGE, Westernblot analysis and N-terminal sequencing. The fluorescence of thecleavage product 7-amino-4-methylcoumarin (AMC) is followed in aSpex-Fluorolog fluorimeter at λ_(ex) =380 nm and λ_(em) =460 nm. Whenthe experiments are carried out at temperatures of 12° C., the cleavageof the substrate is linear, and the autocatalytic activity of calpain islow, over a measurement period of 60 min (see Chatterjee et al. 1996,Bioorg. & Med. Chem. Lett., Vol. 6, 1619-1622). The inhibitors and thecalpain substrate are added to the experimental mixture as DMSOsolutions, in which the final concentration of DMSO should not exceed2%.

In a typical experimental mixture, 10 μl of substrate (250 μm finalconcentration) and then 10 μl of μ-calpain (2 μg/ml final concentration,ie. 18 nM) are added to a 1 ml cuvette which contains buffer. Thecalpain-mediated cleavage of the substrate is measured for from 15 to 20min. 10 μl of inhibitor (50 or 100 μM solution in DMSO) are then addedand inhibition of the cleavage is measured for a further 40 min. K_(i)values are determined using the customary equation for reversibleinhibition, ie. K: l(v_(o) /v)-1 [sic]; where I=inhibitor concentration,v_(o) =initial velocity before adding the inhibitor; v_(i) =reactionvelocity at equilibrium.

The K_(I) for(S)-N(1-ethoxycarbonyl-1-oxo-3-phenyl-2-propanyl)-2-phenylbenzamide(Example 24) was found to be <10 μM. This derivative is thereforemarkedly more effective than is the very closely relatedN(1-ethoxycarbonyl-1-oxo-3-phenylpropan-2-yl)-benzamide (from M. R.Angelastro et al., J. Med. Chem. 1990, 33, 11-13).

Calpain is an intracellular cysteine protease. Calpain inhibitors haveto be able to pass through the cell membrane in order to preventdegradation by calpain of intracellular proteins. Some known calpaininhibitors, such as E 64 and leupeptin, are only able to pass throughcell membranes with difficulty and consequently only have a poor effectin cells even though they are good inhibitors of calpain. The aim is tofind compounds which are better able to traverse membranes. Humanplatelets were used for demonstrating the membrane-traversing ability ofcalpain inhibitors.

Calpain-mediated Degradation of Tyrosine Kinase pp60src in Platelets

The tyrosine kinase pp60src was cleaved by calpain after platelets hadbeen activated. This was investigated in detail by Oda et al. in J.Biol. Chem., 1993, Vol 268, 12603-12608. This study demonstrated thatthe cleavage of pp60src can be prevented by calpeptin, which is aninhibitor of calpain. The cellular efficacy of the novel substances wastested in accordance with this publication. Fresh human, citrate-treatedblood was centrifuged at 200 g for 15 min. The platelet-rich plasma waspooled and diluted 1:1 with platelet buffer (platelet buffer: 68 mMNaCl, 2.7 mM KCl, 0.5 mM MgCl₂ ×6 H₂ O, 0.24 mM NaH₂ PO₄ ×H₂ O, 12 mMNaHCO₃, 5.6 mM glucose, 1 mM EDTA, pH 7.4). After a centrifugation andwashing step using platelet buffer, the platelets were adjusted to 10⁷cells/ml. The human platelets were isolated at RT.

In the test mixture, isolated platelets (2×10⁶) were preincubated, at37° C. for 5 min, with different concentrations of inhibitors (dissolvedin DMSO). The platelets were then activated with 1 μM ionophore A23187and 5 mM CaCl₂. After an incubation of 5 min, the platelets werecentrifuged briefly at 13,000 rpm and the pellet was taken up in SDSsample buffer (SDS sample buffer: 20 mM tris-HCl, 5 mM EDTA, 5 mM EGTA,1 mM DTT, 0.5 mM PMSF, 5 μg/ml leupeptin, 10 μm pepstatin, 10% glyceroland 1% SDS). The proteins were fractionated in a 12% gel, and pp60srcand its 52 kDa and 47 kDa cleavage products were identified by Westernblotting. The polyclonal rabbit anti-Cys-src (pp60^(c-src)) antibodyemployed was obtained from Biomol Feinchemikalien (Hamburg). Thisprimary antibody was detected using a goat HRP-coupled second antibody(Boehringer Mannheim, FRG). The Western blotting was carried out inaccordance with known methods.

The cleavage of pp60src was quantified densitometrically, with use beingmade, as controls, of non-activated platelets (control 1: no cleavage)and platelets which were treated with ionophore and calcium (control 2:corresponds to 100% cleavage). The ED₅₀ value corresponds to theconcentration of inhibitor at which the intensity of the color reactionof the 60 kDa band corresponds to the value: intensity of control 1 pluscontrol 2 divided by 2.

Glutamate-induced Cell Death in Cortical Neurones

The test was carried out as described in Choi D. W., Maulucci-Gedde M.A. and Kriegstein A. R., "Glutamate neurotoxicity in cortical cellculture". J. Neurosci. 1989, 7, 357-368.

Cortex halves are dissected out from 15-day mouse embryos and theindividual cells are isolated enzymically (trypsin). These cells (gliaand cortical neurones) are sown in 24-well plates. After three days(laminin-coated plates) or seven days (ornithine-coated plates), mitosistreatment is carried out using FDU (5-fluoro-2-deoxyuridine). At 15 daysafter preparing the cells, cell death is induced by adding glutamate (15minutes). The calpain inhibitors are then added after removing theglutamate. 24 hours later, cell damage is ascertained by determininglactate dehydrogenase (LDH) in the cell culture supernatant.

It is postulated that calpain also plays a role in apoptotic cell death(M. K. T. Squier et al. J. Cell. Physiol. 1994, 159, 229-237; T. Patelet al. Faseb Journal 1996, 590, 587-597). For this reason, cell deathwas induced with calcium in the presence of a calcium ionophore inanother model, ie. a human cell line. Calpain inhibitors have to getinto the cell, and there inhibit calpain, in order to prevent the celldeath which has been induced.

Calcium-mediated Cell Death in NT2 Cells

In the human cell line NT2, cell death is induced by calcium in thepresence of the ionophore A 23187. 20 hours before the experiment, cellsare plated out in microtiter plates at the rate of 10⁵ cells/well. Oncethe 20 hours have elapsed, the cells are incubated with differentconcentrations of inhibitors in the presence of 2.5 μM ionophore and 5mM calcium. After 5 hours, 0.05 ml of XTT (Cell Proliferation Kit II,Boehringer Mannheim) is added to each reaction mixture. The opticaldensity is determined approximately 17 hours later, in accordance withthe manufacturer's instructions, in an SLT EASY READER EAR 400. Theoptical density at which half the cells have died is computed from thetwo measurements without inhibitors, which were incubated [sic] in theabsence and presence of ionophore. The concentration of the inhibitorwhich achieves this half-maximum optical density is the IC₅₀ value.

An increase in glutamate activity, which leads to states of superexcitation or toxic effects in the central nervous system (CNS), occursin a number of neurological diseases or psychic disorders.

Consequently, substances which inhibit glutamate-mediated effects can beused to treat these diseases. Glutamate antagonists, which also include,in particular, NMDA antagonists or their modulators and the AMPAantagonists, are suitable for therapeutic use as agents againstneurodegenerative diseases (Huntington's chorea and Parkinson'sdiseases), neurotoxic disturbances following hypoxia, anoxia orischaemia, as occur following a stroke, or as antiepileptics,antidepressives and anxiolytics (cf. Arzneim. Forschung 1990, 40,511-514; TIPS, 1990, 11, 334-338 and Drugs of the Future 1989, 14 (11),1059-1071).

Intracerebral administration of excitatory amino acids (EAA) induces asuperexcitation which is so massive that it rapidly leads to convulsionsand the death of the animals. These symptoms can be inhibited by thesystemic, eg. intraperitoneal, administration of centrally acting EAAantagonists. Since excessive activation of EAA receptors in the centralnervous system plays an important role in the pathogenesis of variousneurological diseases, it can be concluded that substances which aredemonstrated to exhibit EAA antagonism in vivo will be useful in thetherapy of CNS diseases of this nature. These diseases include, interalia, focal and global ischaemias, trauma, epilepsies and variousneurodegenerative diseases, such as Huntington's chorea, Parkinson'sdisease, inter alia.

It has already been shown that calpain inhibitors, too, exhibit aprotective effect against EAA-induced cell death in cell cultures (H.Cauer et al., Brain Research 1993, 607, 354-356; Yu Cheg and A. Y. Sun,Neurochem. Res. 1994, 19, 1557-1564). Surprisingly, the calpaininhibitors which are included in this application are effective evenagainst the convulsions which are induced by EAA (eg. NMDA or AMPA) andconsequently point to a therapeutic use in the abovementioned CNSdiseases.

The ketobenzamides I are inhibitors of cysteine derivatives such ascalpain I and/or II and cathepsin B and/or L and may consequently beused for controlling disorders which are associated with increasedactivity of the calpain or cathepsin enzymes. They are therefore usefulfor treating neurodegenerative disorders which occur following ischemia,trauma, subarachnoid hemorrhage and stroke, and which include, inparticular, cerebral stroke and cranial trauma, for treatingneurodegenerative disorders such as multi-infarct dementia, Alzheimer'sdisease and Huntington's disease and, furthermore, for treating damageto the heart following cardiac ischemias, damage to the kidneysfollowing renal ischemias, skeletal muscle damage, muscular dystrophies,damage which arises due to the proliferation of the smooth muscle cells,coronary vasospasms, cerebral vasospasms, cataracts of the eyes andrestenosis of blood vessels following angioplasty. Furthermore, thebenzamides I can be of use for the chemotherapy of tumors and theirmetastases and for treating disorders in which there is an elevatedlevel of interleukin 1, as in inflammations and rheumatic diseases. Inaddition to the customary drug auxiliary substances, the drugpreparations according to the invention also comprise a therapeuticallyeffective quantity of the compounds I.

For local external use, for example in powders, ointments or sprays, theactive compounds can be present in the customary concentrations. As arule, the active compounds are present in a quantity of from 0.001 to 1%by weight, preferably of from 0.01 to 0.1% by weight.

For internal use, the preparations are administered in single doses. Ina single dose, from 0.1 to 100 mg are administered per kg of bodyweight. The preparation can be administered daily in one or more dosagesdepending on the nature and severity of the diseases.

The drug preparations according to the invention comprise, in additionto the active compound, the customary carriers and diluents inaccordance with the desired mode of administration. Pharmaceuticalauxiliary substances, such as ethanol, isopropanol, ethoxylated castoroil, ethoxylated hydrogenated castor oil, polyacrylic acid, polyethyleneglycol, polyethylene glycostearate, ethoxylated fatty alcohols, paraffinoil, yellow soft paraffin and wool fat can be used for local externalapplications. Lactose, propylene glycol, ethanol, starch, talc andpolyvinylpyrrolidone, for example, are useful for internal applications.

Furthermore, antioxidants, such as tocopherol and butylatedhydroxyanisole, and also butylated hydroxytoluene, taste-improvingadditives, stabilizers, emulsifiers and glidants can also be present.

These substances which are present in the preparation in addition to theactive compound, and also the substances which are used in producing thepharmaceutical preparations, are toxicologically harmless and compatiblewith the relevant active compound. The drug preparations are produced ina customary manner, for example by mixing the active compound with othercustomary carriers and diluents.

The drug preparations can be administered in various ways, for exampleperorally, parenterally, such as intravenously by infusion,subcutaneously, intraperitoneally and topically. Thus, possiblepreparation forms are tablets, emulsions, infusion solutions, injectionsolutions, pastes, ointments, gels, creams, lotions, powders and sprays.

EXAMPLES Example 1(S)-2-(E-2-(Naphth-2-yl)-ethen-1-yl)-N-(1-(N-(3-morpholino-1-yl-propan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide##STR8## a) Ethyl 2-(2-(E-naphth-2-yl)ethen-1-yl)benzoate

29.7 g (0.13 mol) of 2-vinylnaphthalene, 25 g (0.16 mol) of ethyl2-bromobenzoate, 22.5 ml (0.16 mol) of triethylamine, 0.54 g ofpalladium diacetate and 1.44 g of triphenylphosphine were heated at 100°C. for 20 h in 200 ml of acetonitrile. After that, the whole was pouredonto water and this mixture was extracted several times with ethylacetate. The organic phase was concentrated under reduced pressure andthe residue was purified by chromatography on silica gel. Yield: 34g(71%).

b) 2-(E-2-(Naphth-2-yl)ethen-1-yl)benzoic acid

34 g (112.5 mmol) of the intermediate la were dissolved in 200 ml oftetrahydrofuran, and 9.5 g (168.7 mmol) of 80% strength potassiumhydroxide, dissolved in 150 ml of water, were added to this solution.The whole was refluxed for 10 h.

The reaction mixture was then acidified with concentrated hydrochloricacid and extracted with ethyl acetate. The organic phase was washed withwater, dried and concentrated under reduced pressure. The residue wastreated with a further small quantity of ethyl acetate and filtered offwith suction. Yield 23.8 g (78%).

(S)-O-(tert-Butyl)-N-(1-(N-(3-morpholino-1-ylpropan-1-yl)-carbamoyl-3-phenylpropan-1-ol-2-yl)carbamate

1.6 g (10 mmol) of diethyl cyanophosphate and 1.0 g (10 mmol) oftriethylamine are added consecutively, at -5° C., to 2.95 g (10 mmol) ofO-(tert-butyl)2-(S)-N-(1-carboxy-2-hydroxy-3-phenylpropan-1-ol-2-yl)-carbamate (S. L.Harbeson et al., J. Med. Chem. 1994, 37, 2918-29) and 1.4 g (10 mmol) ofN-(3-aminopropan-1-yl)morpholine in 50 ml of anhydrousdimethylformamide. The whole was stirred at -5° C. for 1 h and then atroom temperature for 16 h. It was then poured onto water and thismixture was extracted with ethyl acetate. The organic phase wasextracted with aqueous citric acid solution. This aqueous phase was thenrendered alkaline with dilute sodium hydroxide solution and extractedwith ethyl acetate. The organic phase was dried and concentrated underreduced pressure, with 2.3 g (55%) of product being obtained.

3-(S)-3-Amino-2-hydroxy-3-phenyl-N-(3-morpholin-1-ylpropan-1-yl)butyramide

2.1 g (5 mmol) of the intermediate 1c were dissolved in 60 ml ofmethylene chloride, and 60 ml of trifluoroacetic acid were added to thissolution. The mixture was stirred at room temperature for 30 min. Afterthat, it was concentrated under reduced pressure and the residue wasdissolved in, and reprecipitated from, methylene chloride/ether. 2.4 gof crude product were obtained.

e)2-(S)-2-(E-2-(Naphth-2-yl)ethen-1-yl)-N-(1-(N-(3-morpholino-1-ylpropan-1-yl)carbamoyl)-1-hydroxy-3-phenylpropan-2-yl)-benzamide

0.65 g (4 mmol) of diethyl cyanophosphate and 0.8 g (8 mmol) oftriethylamine were added consecutively, at -5° C., to 2.4 g (4 mmol) ofthe intermediate 1d and 1.1 g (4 mmol) of the intermediate 1b in 30 mlof anhydrous dimethylformamide. The whole was then stirred at -5° C. for1 h and at room temperature for a further 16 h. After that, 200 ml ofwater were added and this mixture was extracted with diethyl ether.

The aqueous phase was neutralized with dilute sodium hydroxide solutionand then extracted with ethyl acetate. This organic phase was dried andconcentrated under reduced pressure. The residue was recrystallized fromethyl acetate. Yield: 0.8g (35%).

f)2-(S)-2-(E-2-(Naphth-2-yl)ethen-1-yl)-N-(1-(N-(3-morpholino-1-ylpropan-1-yl)carbamoyl)-1-oxo-3-phenylpropan-2-yl)benzamide

0.38 g (2.4 mmol) of pyridine/sulfur trioxide complex, dissolved in 4 mlof dimethyl sulfoxide, was added, at room temperature, to 0.46 g (0.8mmol) of the intermediate 1e and 0.3 g (3.2 mmol) of triethylamine in 8ml of dimethyl sulfoxide. The whole was stirred for 16 h. After that,the mixture was first of all diluted with water and then extracted withmethylene chloride. The organic phase was dried and concentrated underreduced pressure. The residue was treated with ether, with 0.3 g (65%)of product resulting.

1H NMR (CDCl₃): δ=1.7(2H), 2.4(6H), 3.2(1H), 3.5(3H), 3.7(4H), 5.8(1H),6.5(1H), 7.0-8.0(19H) and 8.8(1H) ppm.

Example 2(S)-2-(E-2-Naphth-2-yl)ethen-1-yl)-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide[sic] ##STR9## a)2-(S)-O-(tert-Butyl)-N-(1-carbamoyl-3-phenylpropan-1-ol-2-yl)carbamate

217.7 g (60 mmol) of O-(tert-butyl)2-(S)-N-(1-carboxy-2-hydroxy-3-phenylpropan-1-ol-2-yl)carbamate (S. L.Harbeson et al., J. Med. Chem. 1994, 37, 2918-29) were reacted withethanolic ammonia solution in analogy with Example 1c. Yield: 13.5 g(76%).

b) 3-(S)-3-Amino-2-hydroxy-3-phenylbutyramide

13.4 g (45.5 mmol) of the intermediate compound 2a were reacted inanalogy with Example 1d. 12.3 g (88%) of product were obtained.

c)2-(S)-2-(E-2-(Naphth-2-yl)ethen-1-yl)-N-(1-carbamoyl-1-hydroxy-3-phenylprop-2-yl)benzamide

1.26 g (6.6 mmol) of N'-(3-dimethylaminopropyl)-N-ethylcarbodiimidehydrochloride (EDC), 1.85 g (6 mmol) of the intermediate compound 1b and1.2 g (12 mmol) of N-methylmorpholine were added consecutively, at -5°C., to 1.65 g (6 mmol) of the intermediate compound 2b and 0.81 g (6mmol) of 1-hydroxybenzotriazole (HOBT) in 10 ml of anhydrousdimethylformamide. After that, the whole was stirred at -5° C. for 1 hand then at room temperature for a further 16 h. Water was subsequentlyadded and the precipitate was filtered off with suction. Yield: 1.3 g(48%) of product.

d)(S)-2-(2-(Naphth-2-yl)ethen-1-yl)-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide

0.45 g (1 mmol) of the intermediate compound 2c was oxidized in analogywith Example 1f. Yield: 0.28 g (62%). MS: m/e=458(M⁺).

Example 32-(S)-2-(E-2-(3,4-Dimethoxyphenyl)ethen-1-yl)-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide##STR10## a) Ethyl 2-(E-2-(3,4-dimethoxyphenyl)ethen-1-yl)benzoate

5 g (30.5 mmol) of 3,4-dimethoxystyrene were reacted with ethyl2-bromobenzoate in dimethylformamide at 120° C. in analogy with Example1a. 7.2 g (94%) of product were obtained.

b) 2-(E-2-(3,4-Dimethoxyphenyl)ethen-1-yl)benzoic acid

7 g (22 mmol) of the intermediate 3a were hydrolyzed with 4M sodiumhydroxide solution in analogy with Example 1b. Yield: 6.2 g (98%).

c)2-(S)-2-(2-(3,4-Dimethoxyphenyl)ethen-1-yl)-N-(1-carbamoyl-1-hydroxy-3-phenylpropan-2-yl)benzamide

1.7 g (6 mmol) of the intermediate compound 2b were reacted with thecompound 3b in analogy with Example 2c. Yield: 2.1 g (76%).

d)2-(S)-2-(E-2-(3,4-Dimethoxyphenyl)ethen-1-yl)-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide

0.45 g (1 mmol) of the intermediate compound 3c was oxidized in analogywith Example 1f. 0.28 g (62%) of product was obtained.

MS: m/e=479(M⁺).

Example 4(S)-4-(2-Naphthylamido)methyl-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide##STR11## a) 4-(2-Naphthylamido)methylbenzoic acid

12.6 g (66.2 mmol) of 2-naphthoyl chloride, dissolved in 150 ml oftetrahydrofuran, were added dropwise, at 10° C., to 10 g (66.2 mmol) of4-aminomethylbenzoic acid in 150 ml of pyridine. The whole was thenstirred at room temperature for 16 h. The mixture was concentrated underreduced pressure and the resulting residue was purified bychromatography (mobile solvent: methylene chloride/methanol=10/1), with11.3 g (56%) of product resulting.

b)4-(2-Naphthylamido)methyl-N-(3-(S)-1-carbamoyl-1-hydroxy-3-phenylpropan-2-yl)benzamide

1.2 g (4 mmol) of the intermediate 4a were reacted with3-(S)-3-amino-2-hydroxy-3-phenylbutyramide 2b in analogy with Example2c, with 1.7 g (88%) of product resulting.

c)(S)-4-(2-Naphthylamido)methyl-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide

0.48 g (1 mmol) of the intermediate compound 4b was oxidized in analogywith Example 1f. Yield: 0.31 g (65%).

1H NMR (D₆ -DMSO): δ=2.9(1H), 3.2(1H), 4.5(2H), 5.2(1H), 7.0-8.0(16H),8.2(1H), 8.7(1H) and 9.1(2H) ppm.

Example 5(S)-2-Phenyl-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide##STR12## a)2-Phenyl-N-(3-(S)-1-carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-benzamide

0.8 g (4 mmol) of 2-biphenylcarboxylic acid and 1.2 g (4 mmol) ofintermediate compound 2b were reacted in analogy with Example 2c. Yield:1.2 g (80%).

b) (S)-2-Phenyl-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)-benzamide

0.75 g (2 mmol) of the intermediate compound 5a was oxidized in analogywith Example 1f. Yield: 0.35 g (47%).

1H NMR (D₆ -DMSO): δ=2.8(1H), 3.1(1H), 5.2(1H), 7.0-7.5(14H), 7.9(1H),8.1(1H) and 8.9(1H) ppm.

Example 6(S)-2-(Naphth-2-ylmethyl)-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide##STR13## a)4,4-Dimethyl-2-(2-(naphth-2-ylhydroxymethyl)phenyl)-2-oxazoline

104 ml of a 1.6M butyllithium solution were slowly added dropwise, at-78° C., to 25 g (0.14 mol) of 4,4-dimethyl-2-phenyl-2-oxazoline and 0.1g of triphenylmethane in 400 ml of anhydrous tetrahydrofuran. The wholewas stirred for 1 h. After that, the mixture was allowed to warm to -30°and a solution of 20.3 g (0.13 mol) of 2-naphthaldehyde, dissolved in200 ml of anhydrous tetrahydrofuran, was added dropwise. The mixture wasstirred at from -20 to -30° C. for a further 1 h. The reaction solutionwas then allowed to warm to room temperature and the solvent was removedunder reduced pressure. The residue was added to ice water, with thismixture subsequently being extracted with ether. The organic phase wasdried and concentrated under reduced pressure. The residue was purifiedby chromatography (mobile solvent: n-heptane/acetone=40/3). Yield: 25.3g (54%).

b) 3-Napth-2-ylphthalide

22 g (66 mmol) of the intermediate 6a were boiled under reflux for 2 hin a mixture of 250 ml of ethanol and 100 ml of 1M hydrochloric acid.After that, the ethanol was removed under reduced pressure and theresulting precipitate was filtered off with suction. Yield: 16.4 g(95%).

c) 2-Naphth-2-ylbenzoic acid

16 g (61.5 mmol) of the intermediate 6b were dissolved in a mixture of100 ml of tetrahydrofuran and 250 ml of ethanol and then hydrogenatedafter 5 g of palladium/barium sulfate had been added. After that, thewhole was filtered and the filtrate was concentrated under reducedpressure. The residue was recrystallized in toluene, with 13.6 g (85%)of product resulting.

d)2-(Naphth-2-yl)methyl-N-(-3-(S)-1-carbamoyl-1-hydroxy-3-phenylpropan-2-yl)benzamide

1.05 g (4 mmol) of the intermediate 6c were reacted with theintermediate 2b in analogy with Example 2c, with 1.7 g (97%) of theproduct resulting.

e)(S)-2-(Naphth-2-yl)methyl-N-(1-carbamoyl--oxo-3-phenylpropan-2-yl)benzamide

0.88 g (2 mmol) of the intermediate compound 6d was oxidized in analogywith Example 1f. Yield: 0.52 g (60%).

1H NMR (D₆ -DMSO): δ=2.8(1H), 3.2(1H), 4.1(2H), 5.3(1H), 7.1-8.0(17H),8.1(1H) and 8.9(1H) ppm.

Example 7(S)-3-(2-Naphthyl)sulfonamido-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide##STR14## a) Ethyl 3-(2-naphthylsulfonamido)benzoate

34.3 g (0.15 mol) of 2-napthalenesulfonyl chloride, dissolved in 250 mlof tetrahydrofuran, are added dropwise, at 0° C., to 25 g (0.15 mol) ofethyl 3-aminobenzoate and 63 ml (0.45 mol) of triethylamine in 400 ml oftetrahydrofuran. After that, the whole is refluxed for 1 h. The organicsolvent was removed under reduced pressure and the residue waspartitioned between ethyl acetate and water. The ethyl acetate phase wasdried and concentrated under reduced pressure. Yield: 55 g (100%).

b) 3-(2-Naphthylsulfonamido)benzoic acid

55 g (0.15 mol) of the intermediate compound 7a were dissolved in 400 mlof tetrahydrofuran, and 400 ml of 4M sodium hydroxide solution wereadded. The whole was stirred at 60° C. for 1.5 h. The organic solventwas removed under reduced pressure. The remaining aqueous phase wasstirred into dilute hydrochloric acid. The resulting precipitate wasdissolved in ethyl acetate, and this solution was washed with water,dried and concentrated under reduced pressure. The residue was thentreated with methylene chloride. After that, 37.3 g (75%) of productwere obtained.

c)3-(2-Naphthyl)sulfonamido-N-(-3-(S)-1-carbamoyl-1-hydroxy-3-phenylpropan-2-yl)benzamide

0.55 g (1.68 mmol) of the intermediate compound 7b was reacted with thecompound 2b in analogy with example 2c. Yield: 0.72 g (86%).

d)(S)-3-(2-Naphthyl)sulfonamido-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide

0.7 g (1.4 mmol) of the intermediate compound 7c was oxidized in analogywith Example 1f. Yield: 0.68 g (98%).

1H NMR (D₆ -DMSO): δ=2.9(1H), 3.1(1H), 5.2(1H), 7.0-8.1(17H), 8.2(1H),8.8(1H) and 10.5(1H) ppm.

Example 8(S)-3-(2-Naphthyl)sulfonamido-N-(1-N-(3-(imidazol-1-ylpropan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide##STR15## a) Ethyl 3-(S)-3-amino-2-hydroxy-4-phenylbutyrate

28 g (0.12 mol) of 3-(S)-3-amino-2-hydroxy-4-phenylbutyric acid (S. L.Harbeson et al., J. Med. Chem. 1994, 37, 2918-29) were boiled underreflux for 3 h in 500 ml of a 1M ethanolic solution of hydrogenchloride. After that, the whole was concentrated under reduced pressureand the residue was partitioned between water and ethyl acetate. Theethyl acetate phase was rendered alkaline with aqueous sodium hydrogencarbonate solution, in association with which an oil was separated out.This oil was taken up in ethyl acetate and this solution was dried andconcentrated under reduced pressure. Yield: 18 g.

b)3-(Naphth-2-yl)sulfonamido-N-(2-(S)-1-ethoxycarbonyl-1-hydroxy-3-phenylpropan-2-yl)benzamide

16.5 g (50.4 mmol) of the intermediate compound 7b and 11.2 g (50.4mmol) of the compound 8a were reacted in analogy with Example 2c. Yield:7.8 g (30%).

c)3-(2-Naphthyl)sulfonamido-N-(2-(S)-1-carboxy-1-hydroxy-3-phenylpropan-2-yl)benzamide

7.8 g (14.6 mmol) of the intermediate compound 8b were dissolved in 150ml of tetrahydrofuran, and 1.1 g (44 mmol) of lithium hydroxide,dissolved in 20 ml of water, were added. The whole was stirred at roomtemperature for 1 h. After that, the organic solvent was removed underreduced pressure and the aqueous phase was rendered weakly acidic using1M hydrochloric acid. The resulting precipitate was filtered off withsuction. Yield: 7.2 g (98%).

d)3-(Naphth-2-yl)sulfonamido-N-(2-(S)-1-N-(3-(imidazol-1)-ylpropan-1-yl)carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-benzamide

1 g (2 mmol) of the intermediate compound 8c was reacted with3-aminopropan-1-yl-1-imidazole in analogy with Example 2c. Yield: 0.63 g(53%).

e)(S)-3-(2-Naphthyl)sulfonamido-N-(1-N-(3-(imidazol-1-ylpropan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide

0.6 g (0.98 mmol) of the intermediate compound 8d was oxidized inanalogy with Example 1f, with 0.55 g (92%) of product resulting.

Example 9(S)-N-(1-N-(N-Benzylpiperidin-4-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-3-(naphth-2-yl)sulfonamido)benzamide##STR16## a)N-(2-(S)-1-N-(N-Benzylpiperidin-4-yl)carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-3-(naphth-2-yl)sulfonamidobenzamide

1 g (2 mmol) of the intermediate compound 8c and4-amino-N-benzylpiperidine were reacted in analogy with Example 2c, with0.67 g (50%) of product resulting.

b)(S)-N-(1-N-(N-Benzylpiperidin-4-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-3-(naphth-2-yl)sulfonamido)benzamide

0.65 g (1 mmol) of the intermediate compound 9a was oxidized in analogywith Example 1f, with 0.59 g (91%) of product resulting.

Example 10 (S)-2-(E-2-(3,4-Dimethoxyphenyl)ethen-1-yl)-N-(1-N-(3-morpholino-1-ylpropan-1-yl)carbamoyl)-1-oxo-3-phenylpropan-2-yl)benzamide##STR17## a)2-(E-2-(3,4-Dimethoxyphenyl)ethen-1-yl)-N-(2-(S)-1-N-(3-morpholino-1-ylpropan-1-yl)carbamoyl)-1-hydroxy-3-phenylpropan-2-yl)benzamide

1.7 g (6 mmol) of the intermediate compound 3b were reacted with thecompound 2b in analogy with Example 2c. Yield: 1.2 g (34%).

b)(S)-2-(E-2-(3,4-Dimethoxyphenyl)ethen-1-yl)-N-(1-N-(3-morpholino-1-ylpropan-1-yl)carbamoyl)-1-oxo-3-phenylpropan-2-yl)benzamide

0.6 g (1 mmol) of the intermediate compound was oxidized in analogy withExample 1f. Yield: 0.12 g (20%).

1H NMR (CDCl₃): δ=1.8(2H), 2.4-2.7(6H), 3.1(1H), 3.5(2H), 3.6-3.8(5H),3.9(6H), 5.7(1H), 6.3(1H), 6.8-7.9(14H) and 8.5(1H) ppm.

Example 11(S)-3-(Naphth-2-yl)sulfonamido)-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide##STR18## a) 8-Quinolyl-N-(3-ethoxycarbonyl)sulfonamide

5 g (30.3 mmol) of ethyl 3-aminobenzoate were reacted, at 0° C., with8-quinolinesulfonyl chloride in analogy with Example 7a, with 5.9 g(76%) of product resulting.

b) N-(3-Carboxy)-8-quinolylsulfonamide

5.9 g of the intermediate compound 11a were hydrolyzed in analogy withExample 1b. Yield: 5.1 g (95%).

c)3-(Naphth-2-yl)sulfonamido)-N-(3-(S)-1-carbamoyl-1-hydroxy-3-phenylpropan-2-yl)benzamide

1 g (3 mmol) of the intermediate compound 2b were reacted with 0.95 g (3mmol) of the compound 11b in analogy with Example 2c, with 1.3 g (87%)of product resulting.

d)(S)-3-(2-Naphthyl)sulfonamido)-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide

1.2 g (2.4 mmol) of the intermediate compound 11c were oxidized inanalogy with Example 1f. Yield: 0.8 g (70%).

1H NMR (D₆ -DMSO): δ=2.9(1H), 3.1(1H), 5.2(1H), 7.0-8.8(17), 8.1(1H) and10.2(1H) ppm.

Example 12(S)-4-(2-Bromophenylsulfonamido)methyl-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide##STR19## a) O-(tert-Butyl) N-(4-ethoxycarbonylbenzyl)carbamate

7 g (34.7 mmol) of ethyl 4-aminomethylbenzoate and 9.6 ml (39.4 mmol) oftriethylamine were dissolved in 150 ml oftetrahydrofuran/dimethylformamide (2:1), and a solution of 8 g (36.5mmol) of BOC anhydride in 100 ml of tetrahydrofuranwas added dropwise at0° C. The whole was stirred at room temperature for 16 h. The mixturewas then concentrated under reduced pressure and the residue waspartitioned between water and ethyl acetate. The organic phase was driedand concentrated under reduced pressure, with 8.5 g (93%) of productresulting.

b) O-(tert-Butyl) N-(4-carboxybenzyl)carbamate

8.3 g (31.3 mmol) of the intermediate compound 12a were hydrolyzed inanalogy with Example 8c, with 7.3 g (93%) of product resulting.

c)4-(tert-Butyloxyamido)methyl-N-(-3-(S)-1-carbamoyl-1-hydroxy-3-phenylpropan-2-yl)benzamide

7 g (27.9 mmol) of the intermediate compound 12b were reacted with thecompound 2b in analogy with Example 2c. Yield: 9.2 g (77%).

d)4-Aminomethyl-N-(2-(S)-1-carbamoyl-1-hydroxy-3-phenylpropan-2-yl)benzamide

9.0 g (21 mmol) of the intermediate compound 12c were cleaved withtrifluoroacetic acid in analogy with Example 1d. Yield: 10.8 g (100%).

e)4-(Bromophenylsulfonamido)methyl-N-(2-(S)-1-carbamoyl-1-hydroxy-3-phenylpropan-2-yl)benzamide

1.5 g (3.4 mmol) of the intermediate compound 12d were reacted with2-bromobenzenesulfonyl chloride at 0° C. in analogy with Example 7a,with 1.2 g (69%) of product resulting.

f)(S)-4-(2-Bromophenylsulfonamido)methyl-N-(1-carbamoyl-1-oxo-3-phenylpropan-2-yl)benzamide

1.05 g (1.9 mmol) of the intermediate compound 12e were oxidized inanalogy with Example 1f. Yield: 0.78 g (75%).

1H NMR (D₆ -DMSO): δ=2.9(1H), 3.2(1H), 4.2(2H), 5.3(1H), 7.0-8.0(15H),8.4(1H) and 8.8(1H) ppm.

Example 13(S)-N-(1-N-(3-Morpholin-1-yl-3-propan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-2-(naphth-2-ylmethyl)benzamide##STR20## a) O-(tert-Butyl)N-(2-(S)-1-(N-3-morpholin-1-ylpropan-1-yl)carbamoyl-2-hydroxy-3-phenylpropan-2-yl)carbamate

19.2 g (65 mmol) of O-(tert-butyl)2-(S)-N-(1-carboxy-2-hydroxy-3-phenylpropan-1-ol-2-yl)carbamate (S. L.Harbeson et al., J. Med. Chem. 1994, 37, 2918-29) were reacted with1-(aminopropan-1-yl)morpholine in analogy with Example 2c, with 23.5 g(85%) of product resulting.

b)3-(S)-3-Amino-2-hydroxy-N-(3-morpholin-1-ylpropan-1-yl)-4-phenylbutyramide

23.3 g (55.3 mmol) of the intermediate compound 13a were cleaved withtrifluoroacetic acid in analogy with Example 1d, resulting in 28 g ofcrude product, which was subjected to further reaction without beingpurified.

c)N-(2-(S)-1-N-(3-Morpholin-1-ylpropan-1-yl)carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-2-(naphth-2-ylmethyl)benzamide

1.57 g (6 mmol) of the intermediate compound 6c were reacted with thecompound 13b in analogy with Example 2c. Yield: 1.1 g (32%).

d)(S)-N-(1-N-(3-Morpholin-1-yl-3-propan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-2-(naphth-2-ylmethyl)benzamide

0.57 g (1 mmol) of the intermediate compound 13c was oxidized in analogywith Example 2c. Yield: 0.14 g (25%).

1H NMR (D₆ -DMSO): δ=1.6(2H), 2.2(6H), 2.9(1H), 3.2(3H), 3.5(4H),4.1(2H), 5.3(1H), 7.0-7.9(16H) and 8.9(1H) ppm.

Example 14(S)-N-(1-N-(3-Morpholin-1-yl-3-propan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(napth-2-ylamido)methylbenzamide##STR21## a)N-(2-(S)-1-N-(3-Morpholin-1-yl-3-propan-1-yl)carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-4-(napth-2-ylamidomethyl)-benzamide

3.1 g (10 mmol) of the intermediate compound 4a were reacted with thecompound 13b in analogy with Example 2c, with 1.9 g of product beingobtained.

b)(S)-(1-N-(3-Morpholin-1-yl-3-propan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(napth-2-ylamido)methylbenzamide

1.2 g (2 mmol) of the intermediate compound 14a were oxidized in analogywith Example 1f. Yield: 0.83 g (73%).

1H NMR (D₆ -DMSO): δ=1.6(2H), 2.2(6H), 3.0(1H), 3-3.2(3H), 3.5(4H),4.6(2H), 5.2(1H), 6.9-8.0(16H), 8.4(1H), 8.8(1H) and 9.0(1H) ppm.

Example 15(S)-N-(1-N-(3-Morpholin-1-ylpropan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-2-phenylbenzamide##STR22## a) N-(2-(S)-1-N-(3-Morpholin-1-ylpropan-1-yl)carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-2-phenylbenzamide

2 g (10 mmol) of 2-biphenylcarboxylic acid were reacted with theintermediate compound 13b in analogy with Example 2c, with 1.8 g ofproduct being obtained.

b)(S)-N-(1-N-(3-Morpholin-1-ylpropan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-2-phenylbenzamide

1.0 g (2 mmol) of the intermediate compound 15a was oxidized in analogywith Example 1f. Yield: 0.45 g (45%). 1H NMR (D₆ -DMSO): δ=1.7(2H),2.2(6H), 2.8(1H), 3.2(3H),

3.6(4H), 5.2(1H), 7.0-7.8(14H) and 8.9(2H) ppm.

Example 16(S)-2-Methyl-N-(1-N-(3-morpholin-1-ylpropan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-5-(naphth-2-yl-sulfonamido)benzamide##STR23## a) Ethyl 5-amino-2-methylbenzoate

26.5 g (127 mmol) of ethyl 2-methyl-5-nitrobenzoate were hydrogenated inethanol after having added 1 g of palladium/charcoal (10% strength).After filtering, the filtrate was concentrated under reduced pressure.Yield: 0.1 g (89%).

b) Ethyl 2-methyl-5-(naphth-2-ylsulfonamido)benzoate

12.6 g (70.4 mmol) of the intermediate compound 16a were reacted with2-naphthalenesulfonyl chloride at 0° C. in analogy with Example 7a, with20.1 g of product resulting.

c) 2-Methyl-5-(naphth-2-ylsulfonamido)benzoic acid

20 g (54 mmol) of the intermediate compound 16b were hydrolyzed inanalogy with Example 8c, with 15.8 g of product being obtained.

d)2-Methyl-N-(2-(S)-1-N-(3-morpholin-1-ylpropan-1-yl)carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-5-(naphth-2-ylsulfonamido)benzamide

3.4 g (10 mmol) of the intermediate compound 16c were reacted with thecompound 13b in analogy with Example 2c. Yield: 3.8 g.

e)(S)-2-Methyl-N-(1-N-(3-morpholin-1-ylpropan-1-yl)carbamoyl-1-oxo-3-phenylpropan-2-yl)-5-(naphth-2-ylsulfonamido)benzamide

0.92 g (1.5 mmol) of the intermediate compound was oxidized in analogywith Example 1f. Yield: 0.3 g (32%).

1H NMR (D₆ -DMSO): δ=1.6(2H), 2.0(3H), 2.3(3H), 2.8(1H), 3.2(2H),3.2-3.5(3H), 3.6(4H), 5.2(1H), 6.9-8.1(14H), 8.3(1H), 8.7(1H), 8.9(1H)and 10.4(1H) ppm.

Example 17(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-2-methyl-5-(naphth-2-ylsulfonamido)benzamide##STR24## a)N-(2-(S)-1-Carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-2-methyl-5-(naphth-2-ylsulfonamido)benzamide

2.7 g (8 mmol) of the intermediate compound 16c were reacted with thecompound 2b in analogy with Example 2c. Yield: 1.5 g (46%).

b)(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-2-methyl-5-(naphth-2-ylsulfonamido)benzamide1.0 g (2 mmol) of the intermediate compound 17a was oxidized in analogywith Example 1f. Yield: 0.65 g (65%).

1H NMR (D₆ -DMSO): δ=2.0(3H), 2.8(1H), 3.2(1H), 5.2(1H), 6.8-8.0(15H),8.2(2H), 8.6(1H) and 10.2(1H) ppm.

Example 18(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinoxalin-2-ylamido)methylbenzamide##STR25## a)N-(2-(S)-1-Carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-4-(quinoxalin-2-ylamido)methylbenzamide

1.2 g (2.7 mmol) of the intermediate compound 12d were reacted with2-quinoxalinecarbonyl chloride in analogy with Example 7a, with 0.8 g(62%) of product resulting.

b)(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinoxalin-2-ylamido)methylbenzamide

0.78 g (1.6 mmol) of the intermediate compound 18a was oxidized inanalogy with Example 1f. Yield: 0.42 g (55%). MS: m/e=481 (M⁺).

Example 19(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinolin-4-ylamido)methylbenzamide##STR26## a)N-(2-(S)-1-Carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-4-(quinolin-4-ylamido)methylbenzamide

0.8 g (1.8 mmol) of the intermediate compound 12d was reacted with4-quinolinecarboxylic acid in analogy with Example 2c, with 0.4 g (46%)of product resulting.

b)(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinolin-4-ylamido)methylbenzamide

0.39 g (0.8 mmol) of the intermediate compound 19a was oxidized inanalogy with Example 1f. Yield: 0.27 g (70%).

1H NMR (D₆ -DMSO): δ=2.9(1H), 3.1(1H), 4.4(2H), 5.2(1H), 7.0-8.0(15H),8.8(1H), 8.9(1H) and 9.3(2H) ppm.

Example 20(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinoxalin-6-ylamido)methylbenzamide##STR27## a)N-(2-(S)-1-Carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-4-(quinoxalin-6-ylamido)methylbenzamide

0.8 g (1.8 mmol) of the intermediate compound 12d was reacted with6-quinoxalinecarboxylic acid in analogy with Example 2c, with 0.36 g(42%) of product resulting.

b)(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinoxalin-6-ylamido)methylbenzamide

0.35 g (0.72 mmol) of the intermediate compound 20a was oxidized inanalogy with Example 1f. Yield: 0.23 g (66%).

1H NMR (D₆ -DMSO): δ=2.8(1H), 3.2(1H), 4.6(2H), 5.2(1H), 7.0-8.2(10H),8.7(1H), 8.8(1H), 9.0(2H) and 9.4(2H) ppm.

Example 21(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinolin-6-ylamido)methylbenzamide##STR28## a)N-(2-(S)-1-Carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-4-(quinolin-6-ylamido)methylbenzamide

0.8 g (1.8 mmol) of the intermediate compound 12d were reacted with6-quinolinecarboxylic acid in analogy with Example 2c, with 0.41 g (47%)of product resulting.

b)(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinolin-6-ylamido)methylbenzamide

0.4 g (0.83 mmol) of the intermediate compound 21a was oxidized inanalogy with Example 1f. Yield: 0.34 g (85%).

1H NMR (D₆ -DMSO): δ=2.9(1H), 3.1(1H), 4.4(2H), 5.2(1H) and 7.0-9.2(19H)ppm.

Example 22(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinolin-3-ylamido)methylbenzamide##STR29## a)N-(2-(S)-1-Carbamoyl-1-hydroxy-3-phenylpropan-2-yl)-4-(quinoxalin-3-ylamido)methylbenzamide

1.0 g (2.3 mmol) of the intermediate compound 12d were 45 reacted with3-quinoxalinecarboxylic acid in analogy with Example 2c, with 0.89 g(80%) of product resulting.

b)(S)-N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-4-(quinoxalin-6-ylamido)methylbenzamide

0.84 g (1.7 mmol) of the intermediate compound 22a was oxidized inanalogy with Example 1f. Yield: 0.75 g (90%). MS: m/e=480 (M⁺).

Example 23N-(1-Ethoxycarbonyl-1-oxo-3-phenylpropan-2-yl)-4-(naphth-2-ylamido)benzamide##STR30## a) 3-(Naphth-2-ylamido)benzoic acid

14.8 g (0.11 mol) of 3-aminobenzoic acid were dissolved in 300 ml ofpyridine, and 20.6 g (0.11 mol) of 2-naphthoyl chloride were added inportions. The whole was stirred at room temperature for 16 h. Themixture was then concentrated under reduced pressure and the residue wasrecrystallized from ethanol. Yield: 30.3 g (97%).

b) N-(1-Ethoxycarbonyl-3-phenylpropan-2-yl)-4-(naphth-2-ylamido)benzamide

18.0 g (61.8 mmol) of the intermediate compound 23a and 14.2 g (61.8mmol) of D,L-alanine ethyl ester were reacted in analogy with Example2c, with 19.8 g (71%) of product being obtained.

c) N-(1-Carboxy-3-phenylpropan-2-yl)-4-(naphth-2-ylamido)benzamide

19.5 g (41.8 mmol) of the intermediate compound 23b were hydrolyzed inanalogy with Example 8c. Yield: 15.2 g (83%).

d)N-(1-Ethoxycarbonyl-1-oxo-3-phenylpropan-2-yl)-4-(naphth-2-ylamido)benzamide

7.1 ml (63.9 mmol) of ethyl oxalyl chloride were added dropwise to asolution of 14.0 g (32 mmol) of the intermediate compound 23c, 0.4 g(3.2 mmol) of N,N-4-dimethylaminopyridine and 10.3 ml (127.7 mmol) ofpyridine in 100 ml of anhydrous tetrahydrofuran such that thetemperature rose to approx. 40° C. The whole was then boiled underreflux for 3 h. It was then stirred at room temperature for a further 16h. 100 ml of water were then added carefully and the mixture was stirredonce again for 30 min. A large quantity of water was added to thereaction mixture and the whole was extracted with ethyl acetate. Theorganic phase was dried and concentrated under reduced pressure, therebyyielding 17 g of an oil. This oil was dissolved in 100 ml of absoluteethanol and 0.24 g of potassium tert-butoxide was added. The mixture wasstirred at room temperature for a further 16 h. It was then concentratedunder reduced pressure and the residue was purified by is chromatography(mobile solvent: methylene chloride/ethyl acetate=10/1). Yield: 7.5 g(54%).

1H NMR (CDCl₃): δ=1.3(3H), 3.2(1H), 3.3(1H), 4.2(2H), 5.6(1H) and6.9-8.4(18H) ppm.

Example 24(S)-N-(1-Ethoxycarbonyl-1-oxo-3-phenylpropan-2-yl)-2-phenylbenzamide##STR31## a)N-(3-(S)-1-Ethoxycarbonyl-1-hydroxy-3-phenylpropan-2-yl)-2-phenylbenzamide

2-Biphenylcarboxylic acid was reacted with methyl3-(S)-3-amino-2-hydroxy-4-phenylbutyrate in analogy with Example 2c.

b) (S)-N-(1-Ethoxycarbonyl-1-oxo-3-phenylpropan-2-yl)-2-phenylbenzamide

The intermediate compound 24a was oxidized in analogy with Example 1f.MS: m/e=387 (M⁺).

Example 25(S)-N(N-Carboxymethyl-1-carbamoyl-1-oxo-3-phenylpropan-2-yl)-3-(2-naphthylsulfonamido)benzamide##STR32## a)O-tert-Butyl-N(3(S)-1-ethoxycarbonyl-2-hydroxy-4-phenylpropan-2-yl)urethane

2.3 g (7.7 mMol [sic]) ofO-tert-butyl-N-(3(S)-1-carboxy-2-hydroxy-4-phenylpropan-2-yl)urethaneand 1.1 g (7.7 mmol) of glycine ethyl ester hydrochloride were reactedin analogy with Example 2c, with 1.7 g (57%) of the product beingobtained.

b)3(S)-3-Amino-N-(ethoxycarbonylmethyl)-2-hydroxy-4-phenyl-butyramide×trifluoroaceticacid

1.4 g (3.7 mMol [sic]) of the intermediate compound 25a were dissolvedin 25 ml of methylene chloride and this solution was stirred at roomtemperature for 2 h after 10 ml of trifluoroacetic acid had been added.The whole was then concentrated under reduced pressure, with 1.5 g(100%) of the product resulting.

c)(S)-N(1-(N-Ethoxycarbonylmethylcarbamoyl)-1-hydroxy-3-phenylpropan-2-yl)-3-(2-naphthylsulfonamido)benzamide

The intermediate compound 7b was reacted with the product 25b in analogywith Example 2c. Yield: 1.3 g

d)(S)-N(1-(N-Carboxymethylcarbamoyl)-1-hydroxy-3-phenylpropan-2-yl)-3-(2-naphthylsulfonamido)benzamide

1.2 g (2 mMol [sic]) of the intermediate compound 25c were hydrolyzedwith lithium hydroxide in analogy with Example 8c. Yield: 0.77 g (67%).

e)(S)-N(1-(N-Carboxymethylcarbamoyl)-1-oxo-3-phenylpropan-2-yl)-3-(2-naphthylsulfonamido)benzamide

0.7 g (1.2 mMol [sic]) of the intermediate compound 25d were oxidized inanalogy with Example 1f, with 0.16 g (23%) of the product beingobtained. MS: m/e=559 (M⁺).

Example 26N-(1-Carbamoyl-1-oxo-3-phenylpropan-2-yl)-3-(2-naphthylsulfonamido)benzamide##STR33## a) The intermediate compound 7b was reacted with ethyl3-amino-2-hydroxy-4-phenylbutyrate in analogy with Example 2c.

b)N(N-Carboxymethyl-1-carbamoyl-1-oxo-3-phenylpropan-2-yl)-3-(2-naphthylsulfonamido)benzamide

The intermediate compound 26a was oxidized in analogy with Example 1f,with the product being obtained.

¹ H-NMR (D₆ -DMSO): δ=2.5(2H), 5.2(1H), 7.1-8.1(17H), 8.4(2H), 8.8(1H)and 10.5(1H) ppm.

The following can be prepared in an analogous manner:

       -      ##STR34##     Example Stereo-chemistry at C-2 R.sup.1 X R.sup.2 R.sup.3 R.sup.4       27 (S)      ##STR35##      ##STR36##      H CH.sub.2      --Ph NH.sub.2                28 (S)      ##STR37##      ##STR38##      H CH.sub.2      --Ph NH.sub.2                29 (S)      ##STR39##      ##STR40##      H CH.sub.2      --Ph            ##STR41##     30 (S)      ##STR42##      ##STR43##      H CH.sub.2      --Ph            ##STR44##     31 (R,S)      ##STR45##      ##STR46##      H --(CH.sub.2).sub.3      CH.sub.3 NH.sub.2                          32 (S)      ##STR47##      ##STR48##      H CH.sub.2      --Ph NH.sub.2                33 (S)      ##STR49##      ##STR50##      H CH.sub.2 --Ph NHCH.sub.2      CH.sub.3                                34 (R,S)      ##STR51##      ##STR52##      H CH.sub.2 CH.sub.2 CH.sub.2 CH.sub.2      CH.sub.3 NH.sub.2                                           35 (S)      ##STR53##      ##STR54##      H CH.sub.2      Ph            ##STR55##     36 (S)      ##STR56##      ##STR57##      H CH.sub.2      --Ph NH.sub.2                37 (S)      ##STR58##      ##STR59##      H CH.sub.2      --Ph            ##STR60##     38 (S)      ##STR61##      ##STR62##      H CH.sub.2 --Ph NHCH.sub.2 CH.sub.2      CH.sub.3                                         39 (R,S)      ##STR63##      4-SO.sub.2 NHCH.sub.2 H CH.sub.2      --Ph NH.sub.2                                      40 (R,S)      ##STR64##      4-SO.sub.2 NHCH.sub.2 H CH.sub.2 --Ph NHCH.sub.2 CH.sub.2 CH.sub.3                                                                         41     (S)      ##STR65##      4-CONH-- H CH.sub.2      --Ph NH.sub.2                         42 (S)      ##STR66##      3-SO.sub.2 NH 6-CH.sub.3 CH.sub.2      --Ph NH.sub.2                                       43 (S)      ##STR67##      3-SO.sub.2      NH 6-CH.sub.3            ##STR68##      NH.sub.2     44 (R,S)      ##STR69##      3-SO.sub.2 NH 6-CH.sub.3 CH.sub.2 CH.sub.2 CH.sub.2 CH.sub.3 NH.sub.2     45 (S)      ##STR70##      3-SO.sub.2 NH 6-CH.sub.3 CH.sub.2      --Ph            ##STR71##     46 (S)      ##STR72##      3-SO.sub.2 NH 6-CH.sub.3 CH.sub.2      --Ph            ##STR73##     47 (S) Ph-- 3-SO.sub.2 NH H CH.sub.2      --Ph NH.sub.2                        48 (R,S) Ph 3-SO.sub.2      NH 6-CH.sub.3 CH.sub.2      --Ph NH.sub.2                            49 (S) Ph 3-SO.sub.2 NH H CH.sub.2      --Ph            ##STR74##     50 (S) Ph 3-SO.sub.2 NH H CH.sub.2 --Ph NHCH.sub.2 CH.sub.2 CH.sub.3                                                                          51     (S) Ph 3-SO.sub.2 NH H CH.sub.2      --Ph            ##STR75##     52 (R,S) Ph 3-SO.sub.2 NH 6-CH.sub.3 CH.sub.2 CH.sub.2 CH.sub.2 CH.sub.2       CH.sub.3 NH.sub.2       53 (S) Ph 3-SO.sub.2 NH 6-CH.sub.3 CH.sub.2 CH.sub.2 CH.sub.2 CH.sub.2       CH.sub.3 NH.sub.2     54 (R,S)      ##STR76##      ##STR77##      H CH.sub.2 CH.sub.2 CH.sub.2      CH.sub.3 NH.sub.2                                  55 (R,S)      ##STR78##      ##STR79##      H CH.sub.2 CH.sub.2 CH.sub.2      CH.sub.3            ##STR80##     56 (R,S)      ##STR81##      ##STR82##      H CH.sub.2 CH.sub.2 CH.sub.2 CH.sub.3 NHCH.sub.2      CH.sub.3                                                      57 (S)      ##STR83##      ##STR84##      H CH.sub.2      --Ph NH.sub.2                58 (S)      ##STR85##      ##STR86##      H CH.sub.2      --Ph            ##STR87##     59 (S)      ##STR88##      2-CH.sub.2 --O-- H CH.sub.2      --Ph NH.sub.2                                 60 (S)      ##STR89##      2-CH.sub.2 H CH.sub.2      --Ph NH.sub.2                           61 (R,S)      ##STR90##      2-CH.sub.2 H CH.sub.2 CH.sub.2 CH.sub.2      CH.sub.3 NH.sub.2                                             62 (R,S)      ##STR91##      2-CH.sub.2 H CH.sub.2      Ph            ##STR92##     63 (R,S)      ##STR93##      ##STR94##      H CH.sub.2      Ph NH.sub.2                64 (R,S)      ##STR95##      ##STR96##      H CH.sub.2      Ph NH.sub.2                65 (R,S)      ##STR97##      ##STR98##      H CH.sub.2      Ph NH.sub.2                66 (R,S)      ##STR99##      ##STR100##      H CH.sub.2      Ph NH.sub.2                67 (R,S)      ##STR101##      ##STR102##      H CH.sub.2      Ph            ##STR103##     68 (R,S)      ##STR104##      ##STR105##      H CH.sub.2      Ph NH.sub.2                69 (R,S)      ##STR106##      2-.tbd.-- H CH.sub.2      Ph NH.sub.2                          70 (R,S)      ##STR107##      2-.tbd.-- H CH.sub.2      Ph            ##STR108##     71 (R,S)      ##STR109##      3-SO.sub.2 NH H CH.sub.2      Ph            ##STR110##     72 (R,S)      ##STR111##      3-SO.sub.2 NH H CH.sub.2      Ph            ##STR112##     73 (R,S)      ##STR113##      3-SO.sub.2 NH H CH.sub.2      Ph            ##STR114##     74 (R,S)      ##STR115##      3-SO.sub.2 NH H CH.sub.2      Ph            ##STR116##     75 (R,S)      ##STR117##      ##STR118##      5-NO.sub.2 CH.sub.2      Ph NH.sub.2                         76 (R,S)      ##STR119##      ##STR120##      5-NO.sub.2 CH.sub.2 Ph NHCH.sub.2 CH.sub.2      CH.sub.3                                                77 (R,S)      ##STR121##      ##STR122##      5-NHCOCH.sub.3 CH.sub.2      Ph NH.sub.2                             78 (R,S)      ##STR123##      ##STR124##      5-NHCOPh CH.sub.2      Ph NH.sub.2                       79 (R,S)      ##STR125##      5-SO.sub.2      NH--            ##STR126##      CH.sub.2      Ph NH.sub.2             80 (R,S)      ##STR127##      5-SO.sub.2      NH--            ##STR128##      CH.sub.2      Ph            ##STR129##     81 (R,S)      ##STR130##      4-SCH.sub.2 -- H CH.sub.2      Ph NH.sub.2                               82 (R,S)      ##STR131##      4-SCH.sub.2 -- H CH.sub.2      Ph            ##STR132##     83 (R,S)      ##STR133##      4-SCH.sub.2 -- H CH.sub.2      Ph            ##STR134##     84 (R,S)      ##STR135##      4-SO.sub.2 -- H CH.sub.2      Ph            ##STR136##     85 (R,S)      ##STR137##      ##STR138##      H CH.sub.2      Ph            ##STR139##     86 (R,S)      ##STR140##      ##STR141##      H CH.sub.2      Ph            ##STR142##     87 (R,S)      ##STR143##      ##STR144##      H CH.sub.2      Ph            ##STR145##     88 (R,S)      ##STR146##      ##STR147##      H CH.sub.2      Ph            ##STR148##     89 (R,S)      ##STR149##      ##STR150##      H CH.sub.2      Ph            ##STR151##     90 (R,S)      ##STR152##      ##STR153##      H CH.sub.2      Ph            ##STR154##     91 (R,S)      ##STR155##      ##STR156##      H CH.sub.2      Ph            ##STR157##     92 (R,S)      ##STR158##      ##STR159##      H CH.sub.2      Ph            ##STR160##     93 (R,S)      ##STR161##      ##STR162##      H CH.sub.2 CH.sub.2 CH.sub.2      CH.sub.3 NH.sub.2                                  94 (R,S)      ##STR163##      ##STR164##      H CH.sub.2 CH.sub.2 CH.sub.2      CH.sub.3            ##STR165##     95 (R,S)      ##STR166##      ##STR167##      H CH.sub.2      Ph NH.sub.2                96 (R,S)      ##STR168##      ##STR169##      H CH.sub.2      Ph            ##STR170##     97 (R,S)      ##STR171##      ##STR172##      H CH.sub.2      Ph            ##STR173##     98 (R,S)      ##STR174##      ##STR175##      H CH.sub.2      Ph            ##STR176##     99 (R,S)      ##STR177##      ##STR178##      H CH.sub.2      Ph            ##STR179##     100 (R,S)      ##STR180##      ##STR181##      H CH.sub.2      Ph            ##STR182##     101 (R,S)      ##STR183##      ##STR184##      H CH.sub.2      Ph NH.sub.2                102 (R,S)      ##STR185##      ##STR186##      H CH.sub.2      Ph            ##STR187##     103 (R,S)      ##STR188##      ##STR189##      H CH.sub.2      Ph            ##STR190##     104 (R,S)      ##STR191##      ##STR192##      H CH.sub.2      Ph            ##STR193##     105 (R,S)      ##STR194##      ##STR195##      H CH.sub.2      Ph            ##STR196##     106 (R,S)      ##STR197##      ##STR198##      H CH.sub.2      Ph NH.sub.2                107 (R,S)      ##STR199##      ##STR200##      H CH.sub.2      Ph            ##STR201##     108 (R,S)      ##STR202##      ##STR203##      H CH.sub.2      Ph            ##STR204##     109 (R,S)      ##STR205##      ##STR206##      H CH.sub.2      Ph NH.sub.2                110 (R,S)      ##STR207##      ##STR208##      H CH.sub.2      Ph            ##STR209##     111 (R,S)      ##STR210##      ##STR211##      H CH.sub.2      Ph            ##STR212##     112 (R,S)      ##STR213##      ##STR214##      H CH.sub.2      Ph            ##STR215##

We claim:
 1. A ketobenzamide of the formula I ##STR216## and itstautomeric and isomeric forms, and also, where appropriate, itsphysiologically tolerated salts, where the variables have the followingmeanings:R¹ is phenyl, naphthyl, quinolyl, pyridyl, pyrimidyl, pyrazyl,pyridazyl, quinazolyl, quinoxalyl, thienyl, benzothienyl, benzofuryl,benzimidazolyl, furanyl, indolyl, isoquinoline, tetrahydroisoquinolineor tetrahydroquinoline, where the aromatic and hetero-aromatic rings canadditionally be substituted by one, two or three R⁵ radicals, R² ischlorine, bromine, fluorine, C₁ -C₆ -alkyl, C₂ -C₆ -alkenyl, C₂ -C₆-alkynyl, C₁ -C₆ -alkyl-phenyl, C₂ -C₆ -alkenyl-phenyl, C₂ -C₆-alkynyl-phenyl, phenyl, NHCO--C₁ -C₄ -alkyl, --NHCO-phenyl,--NHCO-naphthyl, H₂ N--SO₂ --C₁₋₄ -alkyl--, COOH, --COO--C₁₋₄ -alkyl,--CONH--C₁₋₄ -alkyl, C₁₋₄ -alkoxy, NO₂ or NH₂, R³ is C₁ -C₆ -alkyl whichcan also carry a phenyl, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, indolyl, pyridyl or naphthyl ring which, forits part, can be substituted by one or two R⁵ radicals, X is a bond,--(CH₂)_(m) --, --(CH₂)_(m) --O--(CH₂)_(o) --, --(CH₂)_(n)--S--(CH₂)_(m) --, --(CH₂)_(n) --SO--(CH₂)_(m) --, --(CH₂)_(n) --SO₂--(CH₂)_(m) --, --CH═CH--, --C.tbd.C--, --CO--CH═CH--, CO--(CH₂)_(m) --,--(CH₂)_(m) --NHCO--(CH₂)_(o) --, --(CH₂)_(m) --CONH--(CH₂)_(o) --,--(CH₂)_(m) --NHSO₂ --(CH₂)_(o) --, --NH--CO--CH═CH--,--CH═CH--CO--NH--, --(CH₂)_(m) --SO₂ NH--(CH₂)_(o) -- or ##STR217## R⁴is OR⁶, NR⁷ R⁸, ##STR218## R⁵ is hydrogen, C₁ -C₄ -alkyl, --O--C₁ -C₄-alkyl, OH, Cl, F, Br, I, CF₃, NO₂, NH₂, CN, COOH, COO--C₁ -C₄ -alkyl,--NHCO--C₁ -C₄ -alkyl, --NHCO-phenyl, --NHSO₂ --C₁ -C₄ -alkyl, --NHSO₂-phenyl, --SO₂ --C₁ -C₄ -alkyl or --SO₂ -phenyl, R⁶ is hydrogen or C₁-C₆ -alkyl which can be substituted by a phenyl ring which, itself, canalso be substituted by one or two R⁹ radicals, R⁷ is hydrogen or C₁ -C₆-alkyl, R⁸ is hydrogen or C₁ -C₆ -alkyl which can also be substituted bya phenyl ring, which can carry one or two R⁹ radicals, or by one of theradicals ##STR219## R⁹ is hydrogen, C₁ -C₄ -alkyl, --O--C₁ -C₄ -alkyl,OH, Cl, F, Br, I, CF₃, NO₂, NH₂, CN, COOH, COO--C₁ -C₄ -alkyl,--NHCO--C₁ -C₄ -alkyl, --NHCO-phenyl, --NHSO₂ --C₁ -C₄ -alkyl, --NHSO₂-phenyl, --SO₂ --C₁ -C₄ -alkyl or --SO₂ -phenyl, R¹⁰ is hydrogen or C₁-C₆ -alkyl which can be substituted by a phenyl ring which can also besubstituted by one or two R⁹ radicals, n is the number 0, 1 or 2, m isthe number 0, 1, 2, 3 or 4, and o is the number 0, 1, 2, 3 or
 4. 2. Aketobenzamide of the formula I as claimed in claim 1, whereR² ishydrogen, C₁ -C₄ -alkyl, fluorine or chlorine, R³ is --CH₂ --phenyl,--CH₂ -cyclohexyl, n-butanyl or n-pentanyl, each of which can besubstituted by an R⁵ radical, R⁴ is --NR⁸, and R¹, X and n have themeanings given in claim
 1. 3. A method of controlling disorders mediatedby calpain enzymes comprising administering to a host in need thereof aneffective amount of a ketobenzamide of the formula I as claimed inclaim
 1. 4. A drug preparation which comprises at least oneketobenzamide of the formula I as claimed in claim 1.